Ethanol production by microorganisms

A kind of ethanol and biological technology, which is applied in the direction of microorganisms, microorganisms, and methods based on microorganisms, and can solve problems such as inefficiency, laboriousness, and time-consuming

Inactive Publication Date: 2011-10-12
JOULE UNLTD TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, genetic manipulation techniques (knockout, overexpression of transgenes via integration or episomic plasmid amplification) are inefficient, time-consuming, laborious or non-existent

Method used

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  • Ethanol production by microorganisms
  • Ethanol production by microorganisms
  • Ethanol production by microorganisms

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Experimental program
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Embodiment 20

[0370] In Example 20, a method for preparing an isoprenoid compound using one or more microorganisms is described. In certain embodiments, the fuel composition prepares 3-methyl-3-buten-1-ol from 3-methyl-3-buten-1-ol by utilizing one or more microorganisms to produce 3-methyl-3-buten-1-ol. Methyl-1-butanol, and the addition of 3-methyl-1-butanol to the fuel composition produces. In other embodiments, the fuel composition prepares 3-methyl-2-buten-1-ol from 3-methyl-2-buten-1-ol by utilizing one or more microorganisms to produce 3-methyl-2-buten-1-ol. Methyl-1-butanol, and the addition of 3-methyl-1-butanol to the fuel composition produces.

[0371] Impurities

[0372] In certain aspects, the carbon-based products (e.g., ethanol, fatty acids, paraffins, isoprenoids) produced herein comprise a mixture that is typically associated with biofuels derived from triglycerides, such as fuels derived from vegetable oils and fats. than less impurities. For example, crude fatty acid ...

Embodiment 1

[0436] Plasmid Construction of Synechococcus sp. PCC 7002

[0437] Construction of pJB5: The pJB5 base plasmid was designed as an empty expression vector for recombination into Synechococcus sp. PCC7002. Two regions of homology, an upstream region of homology (UHR) and a downstream region of homology, were designed to flank the construct. These 500bp homologous regions correspond to positions 3301-3800 and 3801-4300 of UHR and DHR, respectively (Genbank accession number: NC_005025). The aadA promoter, gene sequence and terminator were designed to provide resistance to spectinomycin and streptomycin to the integrated construct. For example, pJB5 was designed with the aph2 kanamycin resistance cassette promoter and ribosome binding site (RBS). Downstream of this promoter and RBS, we designed and inserted restriction endonuclease recognition sites for NdeI and EcoRI and sites for XhoI, BamHI, SpeI and PacI. After the EcoRI site, the native terminator from the alcohol dehydroge...

Embodiment 2

[0445] Plasmid construction for Thermosynechococcus elongates BP-1

[0446] Thermosynechococcus elongates BP-1 was selected as another typical carbon dioxide fixation production host, and was modified by engineered nucleic acid to lose some genes and / or express or overexpress some genes.

[0447] Four plasmids (pJB18, pJB19, pJB20 and pJB21 ), all derivatives of pJB5, were constructed to enable homologous recombination into four different sites of the Synechococcus elongatus BP-1 genome. Specifically, the 0.5 kb upstream homology (UH) and downstream homology (DH) regions used for homologous recombination in Synechococcus PCC 7002 species in pJB5 were replaced by the following approximately 2.5 kb T. elongatus BP-1 (accession number NC_004113) Region substitutions: for pJB18, fit 831908-834231 (UH) and 834232-836607 (DH) genome; for pJB19, fit 454847-457252 (UH) and 457252-459740 (DH); for pJB20, fit 481310-483712 (UH) and 483709-486109 (DH); and for pJB21, fit 787356-789654 (...

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Abstract

The present disclosure relates to methods and compositions for engineering photo autotrophic organisms to convert carbon dioxide and light into fatty acid esters and other molecules, including biofuels. The molecules are then secreted by the organism into a growth medium.

Description

technical field [0001] The present disclosure relates to methods and compositions for engineering photoautotrophic organisms to convert carbon dioxide and light into fatty acid esters and other molecules that are then secreted by the organism into the culture medium. Background technique [0002] Photosynthesis is the process by which living entities use sunlight and carbon dioxide to produce sugars for energy. Due to natural evolution, photosynthesis is an extremely complex system with numerous and poorly understood feedback loops, control mechanisms and process inefficiencies. This complex system presents potentially insurmountable obstacles to both one-factor-at-a-time and global optimization methods [Nedbal et al., Photosynth Res., 93(1-3):223-34( 2007); Salvucci et al., Physiol Plant., 120(2):179-186 (2004); Greene et al., Biochem J., 404(3):517-24 (2007)]. [0003] Existing photoautotrophic organisms such as plants, algae, and photosynthetic bacteria are poorly adapt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/06C12N1/21
CPCC12N1/20C12N9/00C12N9/0006C12N9/0008C12N9/001C12N9/0083C12N9/1007C12N9/1029C12N9/88C12P1/04C12P5/00C12P7/00C12P7/6409C12P7/6427C12P7/6436C12P7/6463C12P7/6472C12P7/649C12P13/04C12Y103/03006C12P5/02C12P5/026C12P7/065Y02E50/10Y02E50/30C12R2001/01C12N1/205C12P7/6458C12P7/6434C12N15/74
Inventor B·格林N·雷帕斯D·罗伯特森
Owner JOULE UNLTD TECH
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